3型BRET，一种改进的基于竞争的生物发光共振能量转移检测方法。
Type-3 BRET, an improved competition-based bioluminescence resonance energy transfer assay.
作者信息
Felce James H, Knox Rachel G, Davis Simon J
机构信息
Radcliffe Department of Clinical Medicine and Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.
Radcliffe Department of Clinical Medicine and Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.
出版信息
Biophys J. 2014 Jun 17;106(12):L41-3. doi: 10.1016/j.bpj.2014.04.061.
Abstract
We show that in conventional, competition-based bioluminescence resonance energy transfer (BRET) assays of membrane protein stoichiometry, the presence of competitors can alter tagged-protein density and artifactually reduce energy transfer efficiency. A well-characterized monomeric type I membrane protein, CD86, and two G protein-coupled receptors β2AR and mCannR2, all of which behave as dimers in these conventional assays, exhibit monomeric behavior in an improved competition-based type-3 BRET assay designed to circumvent such artifacts.
摘要
我们表明，在基于竞争的传统生物发光共振能量转移（BRET）膜蛋白化学计量分析中，竞争者的存在会改变标记蛋白的密度，并人为降低能量转移效率。一种特征明确的单体I型膜蛋白CD86，以及两种G蛋白偶联受体β2AR和mCannR2，在这些传统分析中均表现为二聚体，但在旨在规避此类假象而改进的基于竞争的3型BRET分析中表现出单体行为。
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2
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3
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